Expression, purification, crystallization and preliminary X-ray diffraction analysis of Bifidobacterium adolescentis xylose isomerase

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 May 1;69(Pt 5):588-91. doi: 10.1107/S174430911301110X. Epub 2013 Apr 30.

Abstract

Xylose isomerase (EC 5.3.1.5) is a key enzyme in xylose metabolism which is industrially important for the transformation of glucose and xylose into fructose and xylulose, respectively. The Bifidobacterium adolescentis xylA gene (NC_008618.1) encoding xylose isomerase (XI) was cloned and the enzyme was overexpressed in Escherichia coli. Purified recombinant XI was crystallized using the sitting-drop vapour-diffusion method with polyethylene glycol 3350 as the precipitating agent. A complete native data set was collected to 1.7 Å resolution using a synchrotron-radiation source. The crystals belonged to the orthorhombic space group P21212, with unit-cell parameters a = 88.78, b = 123.98, c = 78.63 Å.

Keywords: Bifidobacterium adolescentis; bioethanol; xylose isomerases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldose-Ketose Isomerases / chemistry
  • Aldose-Ketose Isomerases / genetics*
  • Aldose-Ketose Isomerases / isolation & purification*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / isolation & purification*
  • Bifidobacterium / enzymology*
  • Bifidobacterium / genetics
  • Crystallization
  • Gene Expression Regulation, Bacterial*
  • X-Ray Diffraction

Substances

  • Bacterial Proteins
  • Aldose-Ketose Isomerases
  • xylose isomerase